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|Sample type||Average %||Range|
|Serum||99||89% - 116%|
|Cell culture media||106||103% - 110%|
|Fetal Bovine Serum||111||108% - 115%|
PCNA in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit (ab196270) is designed for the quantitative measurement of PCNA protein in Human cell and tissue extracts.
The SimpleStep ELISA™ employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.
Proliferating cell nuclear antigen (PCNA) is a 28.7 kDa protein which is an essential component of DNA replication. PCNA protein forms a homotrimeric ring structure which assembles around DNA forming a processivity sliding clamp. Levels of PCNA protein are low in quiescent cells and increase upon mitogen stimulation with peak protein levels localized to the nucleus during S-phase. PCNA interacts with a large number of proteins involved in DNA replication, cell cycle control, and cell cycle check point control.
|Components||1 x 96 tests|
|10X Human PCNA Capture Antibody||1 x 600µl|
|10X Human PCNA Detector Antibody||1 x 600µl|
|10X Wash Buffer PT (ab206977)||1 x 20ml|
|50X Cell Extraction Enhancer Solution (ab193971)||1 x 1ml|
|5X Cell Extraction Buffer PTR (ab193970)||1 x 10ml|
|ab221825 - Antibody Diluent 5BI||1 x 6ml|
|Human PCNA Lyophilized Recombinant Protein||2 vials|
|Plate Seals||1 unit|
|Sample Diluent NS||1 x 12ml|
|SimpleStep Pre-Coated 96-Well Microplate (ab206978)||1 unit|
|Stop Solution||1 x 12ml|
|TMB Substrate||1 x 12ml|
Our Abpromise guarantee covers the use of ab196270 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Sandwich ELISA||Use at an assay dependent concentration.|
Background-subtracted data values (mean +/- SD) are graphed.
Background-subtracted data values (mean +/- SD, n = 2) are graphed.
HeLa cells were untreated asynchronous culture and 10 µM Etoposide, 24 hr (S/G2 arrest). PCNA levels were expected to peak during DNA replication in S-phase and return back to normal shortly after exiting S-phase. Background-subtracted data values (mean +/- SD, n = 2) are graphed.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"